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目的探讨生物素对高糖损伤胰岛β细胞功能的影响。方法采用Dextran密度梯度离心法分离纯化大鼠胰岛,在含5.5mmol·L-1葡萄糖、5.5mmol·L-1葡萄糖+1μmol·L-1生物素、含27.0mmol·L-1葡萄糖及27.0mmol·L-1葡萄糖+1μmol·L-1生物素的RPMI1640培养基中培养48h,分析3.3、27.0mmol·L-1葡萄糖刺激的胰岛素分泌及胰岛细胞中胰岛素的含量;提取总mRNA,RT-PCR扩增前胰岛素源基因,电泳并进行灰度分析。结果在5.5mmol·L-1葡萄糖培养条件下,生物素增加高糖诱导的胰岛素释放及胰岛细胞的胰岛素含量,前胰岛素源mRNA也增加;在27.0mmol·L-1葡萄糖培养48h的胰岛中,胰岛细胞中的胰岛素浓度降低,高糖诱导的胰岛素释放及前胰岛素源mRNA显著减少;生物素的共培养,部分提高了高糖诱导的胰岛素释放及前胰岛素源mRNA含量。结论生物素通过促进胰岛素的合成显著改善了高糖损伤胰岛β细胞的功能紊乱。
Objective To investigate the effect of biotin on pancreatic β-cell function induced by high glucose. Methods Dextran density gradient centrifugation was used to separate and purify the pancreatic islets in rats. The cells were treated with 5.5 mmol·L-1 glucose, 5.5 mmol·L-1 glucose + 1 μmol·L-1 biotin, 27.0 mmol·L-1 glucose and 27.0 mmol · L-1 glucose + 1μmol·L-1 biotin was cultured in RPMI1640 medium for 48h, the insulin secretion and the content of insulin in islet cells stimulated by glucose (3.3,27.0mmol·L-1) were analyzed. The total mRNA was extracted and RT-PCR The pre-insulin gene was amplified, electrophoresed and subjected to gray-scale analysis. Results Under the condition of 5.5mmol·L-1 glucose, biotin increased the insulin release and the insulin content of insulin induced by high glucose, and the mRNA of proinsulin increased. In the islet incubated with 27.0mmol·L-1 glucose for 48h, The insulin concentration in islet cells decreased, the release of insulin induced by high glucose and the mRNA of proinsulin decreased significantly, and the co-culture of biotin partially increased the release of insulin and the content of proinsulin mRNA induced by high glucose. Conclusion Biotin can significantly improve the dysfunction of pancreatic β-cells induced by high glucose by promoting the synthesis of insulin.